The present invention relates to a redox control and monitoring platform and, more particularly, to a redox control and monitoring platform that is to be used in conjunction with another detector during high throughput screening and drug discovery applications.
A great number of studies have demonstrated the importance of the redox environment in the regulation of a number of cellular functions in both normal and diseased states. Such processes include those that involve redox active proteins and enzymes, free radical damage, and oxidative stress. The redox environment can influence both catalysis and binding affinity. Additionally, transcription of DNA into mRNA, the translation of mRNA into proteins, and the rate of transport of glutamate across the nerve synapses have all been shown to depend on the redox environment. The cellular redox environment has also been implicated in the modulation of more complex cellular events such as proliferation and apotosis, and the specific redox environment can also activate certain drugs. It is apparent that the redox environment of a target molecule or species can have a great affect on the efficacy of a particular drug.
Researchers often use high throughput systems to analyze a large library of compounds that could have a desired activity. An example of a system useful for high throughput screening and assay is shown in U.S. Pat. No. 6,238,869 to Kris, et al, which is hereby incorporated by reference. Such systems allow researchers to quickly test a huge number of compounds and discard those that do not show the desired activity or quality. Such systems are especially useful in the drug discovery process because large scale testing of a series of compounds can be accomplished quickly and relatively cheaply. Only those compounds that show desired activity are tested further. Without high throughput technology, the screening of such a large number of compounds would be virtually impossible.
Current high throughput drug screening discovery processes do not provide for the measurement of the redox environment or the active control of the redox environment of a target. Much of the current high throughput technology relies on spectroscopic, especially fluorescent, methods to provide information about how or if a particular compound is reacting with a target. However, researchers cannot generally make such measurements while actively controlling the redox environment.
Many methods do exist to measure the redox environment. An example of such a system is shown in U.S. Pat. No. 4,963,815 to Hafeman. However, such a system does not provide for active control of the redox environment of the sample. Additionally, the systems do not provide other data other than electrical measurements, such as spectroscopic measurements. The systems also are not adapted for use in a high throughput process.
Electrochemistry and spectroscopy have been combined to perform various studies. Heineman, Spectro-electro-chemistry: Combination of Optical and Electrochemical Techniques for Studies of Redox Chemistry, Anal. Chem. 1978, 50, 390–402; Asanov et al., Heteroenergetics of Bovine Serum Albumin Adsorption from Good Solvents Related to Crystallization Conditions, J. of Colloid and Interface Science 1997, 191, 222–235; Johnson et al., Potential-Dependent Enzymatic Activity in an Enzyme Thin-Layer Cell, Anal. Chem 1982, 54, 1377–1383. The thin-layer spectrochemical methods are often used to characterize the fundamentals of electron transfer between and within redox active enzymes and other biomolecules. The change in redox potential changes the ratio of the redox forms of the enzymes that the spectroscopic technique is generally measuring. However, these methods cannot be generally performed under conditions where assays independent of the redox potential can be performed. When independent assays have been performed, they involved immobilized biomolecules on a surface or within a membrane. It is known that this immobilization can alter the biological activity and the nature and extent of their interaction with proteins or other potential binding partners such as drugs. Additionally, these methods are not generally applicable to a wide variety of systems and sample types.
Accordingly, there is a need for a versatile system that can provide measurement and control of the redox environment that is independent of other assays that can be performed on the sample, especially in high throughput screening.